湖北海棠总黄酮对成骨细胞增殖分化及破骨细胞活性的影响

Effects of Total Flavoniods of the Malus Hupehensis on Proliferation and Differentiation of Osteoblasts and Activity of Osteoclasts

目的:探讨湖北海棠总黄酮及其主要成分根皮苷对大鼠成骨细胞增殖分化及破骨细胞活性的影响。方法:取新生大鼠颅骨,用改良组织块消化法分离成骨细胞。MTT法检测药物对成骨细胞增殖的影响,对硝基苯二钠基质动力学法测定细胞内碱性磷酸酶(AKP)的活性。分离培养大鼠破骨细胞。加药共培后检测抗酒石酸酸性磷酸酶(TRACP)活性,检查骨吸收陷窝变化。结果:湖北海棠总黄酮能明显促进成骨细胞增殖;提高成骨细胞内碱性磷酸酶活性。湖北海棠主要成分根皮苷能提高成骨细胞内碱性磷酸酶活性;降低抗酒石酸酸性磷酸酶活性,减少骨吸收陷窝数目,与对照组比较差异有统计学意义。结论:湖北海棠总黄酮能促进成骨细胞的增殖和分化,其主要成分根皮苷能促进成骨细胞的分化,抑制破骨细胞的活性和嗜骨活性。

Objective：To explore the influence of total flavoniods of the Malus hupehensis and its main component phloridzin on proliferation and differentiation of osteoblasts and activity of osteoclasts in vitro.Methods：The osteoblasts from cranium of newborn SD rats were cultured by improved Ⅱcollagenase method.The proliferation of osteoblasts was measured by MTT,and the activity of alkaline phosphatase（AKP） was observed with p-nitrophenyl phosphate（PNPP） method.Osteoclasts were separated and cultured from rats.The activity of tartrate-resistant acid phosphatase（TRACP） and the resorption pit area on the bone slices formed by osteoclasts were measured after exposure to tested drugs.Results：Malus hupehensis.flavoniods strongly stimulated proliferation of osteoblasts and increase AKP activity significantly.Phloridzin which is main component in Malus hupehensis can promote AKP activity of osteoblasts and TRACP activity of osteoclasts.The area and number of resorption pits were markedly restrained.Which was significantly different to control group（P0.01）.Conclusion：Malus hupehensis flavoniods can induce the proliferation and differentiation of cultured osteoblasts in vitro.Phloridzin can also induce differentiation of cultured osteoblasts,but inhibit the differentiation of osteoclasts and bone resorption activity.